Molecular Neurobiology

, Volume 53, Issue 9, pp 5847–5863 | Cite as

Dehydroascorbic Acid Promotes Cell Death in Neurons Under Oxidative Stress: a Protective Role for Astrocytes

  • Andrea García-Krauss
  • Luciano Ferrada
  • Allisson Astuya
  • Katterine Salazar
  • Pedro Cisternas
  • Fernando Martínez
  • Eder Ramírez
  • Francisco Nualart
Article

Abstract

Ascorbic acid (AA), the reduced form of vitamin C, is incorporated into neurons via the sodium ascorbate co-transporter SVCT2. However, this transporter is not expressed in astrocytes, which take up the oxidized form of vitamin C, dehydroascorbic acid (DHA), via the facilitative hexose transporter GLUT1. Therefore, neuron and astrocyte interactions are thought to mediate vitamin C recycling in the nervous system. Although astrocytes are essential for the antioxidant defense of neurons under oxidative stress, a condition in which a large amount of ROS is generated that may favor the extracellular oxidation of AA and the subsequent neuronal uptake of DHA via GLUT3, potentially increasing oxidative stress in neurons. This study analyzed the effects of oxidative stress and DHA uptake on neuronal cell death in vitro. Different analyses revealed the presence of the DHA transporters GLUT1 and GLUT3 in Neuro2a and HN33.11 cells and in cortical neurons. Kinetic analyses confirmed that all cells analyzed in this study possess functional GLUTs that take up 2-deoxyglucose and DHA. Thus, DHA promotes the death of stressed neuronal cells, which is reversed by incubating the cells with cytochalasin B, an inhibitor of DHA uptake by GLUT1 and GLUT3. Additionally, the presence of glial cells (U87 and astrocytes), which promote DHA recycling, reverses the observed cell death of stressed neurons. Taken together, these results indicate that DHA promotes the death of stressed neurons and that astrocytes are essential for the antioxidative defense of neurons. Thus, the astrocyte-neuron interaction may function as an essential mechanism for vitamin C recycling, participating in the antioxidative defense of the brain.

Keywords

Ascorbic acid Dehydroascorbic acid Neuron Astrocyte Cell death Recycling Oxidative stress 

Abbreviations

AA

Ascorbic acid

DHA

Dehydroascorbic acid

DMEM

Dulbecco’s modified Eagle’s medium

FBS

Fetal bovine serum

DOG

Deoxyglucose

GFAP

Glial fibrillary acidic protein

GLUT

Glucose transporter

GSH

Glutathione

IMDM

Iscove’s modified Dulbecco’s medium

PAGE

Polyacrylamide gel electrophoresis

PBS

Phosphate-buffered saline

ROS

Reactive oxygen species

SDS

Sodium dodecyl sulfate

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Andrea García-Krauss
    • 1
  • Luciano Ferrada
    • 1
  • Allisson Astuya
    • 1
  • Katterine Salazar
    • 1
  • Pedro Cisternas
    • 1
  • Fernando Martínez
    • 1
  • Eder Ramírez
    • 1
  • Francisco Nualart
    • 1
    • 2
  1. 1.Center for Advanced Microscopy (CMA) BIO-BIO, Faculty of Biological SciencesConcepcion UniversityConcepciónChile
  2. 2.Departamento de Biología Celular, Facultad de Ciencias BiológicasUniversidad de ConcepciónConcepciónChile

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